Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226
ISSN: 2231-3354
Received on: 29-09-2011
Revised on: 03-10-2011
Accepted on: 08-10-2011
Shailendra Kumar Singh
Associate Professor, Department of
pharmaceutical sciences, Guru
Jambheswar university of Science and
Technology. Hissar-125001. Haryana.
India.
Soukarya Som
Research Scholar, Department of
pharmaceutical sciences, Guru
Jambheswar university of Science and
Technology. Hissar-125001. Haryana.
India.
Upender Shankhwar
Research Scholar, Department of
pharmaceutical sciences, Guru
Jambheswar university of Science and
Technology. Hissar-125001. Haryana.
India.
For Correspondence
Soukarya som
Address- 17A EAST ROAD,
JADAVPUR
KOLKATA-700032, WEST
BENGAL
MOBILE No. 9034752514
Formulation and optimization of solid dispersion
of Clopidogrel with PEG 6000
Shailendra Kumar Singh, Soukarya Som and Upender Shankhwar
ABSTRACT
Clopidogrel, a non competitive inhibitor of adenosine diphosphate at the platelet
receptors, is an anticoagulant drug practically insoluble in water. In order to improve the aqueous
solubility of drug and its dissolution rate solid dispersions of clopidogrel were prepared with
different proportions of the hydrophilic carrier PEG 6000. A two factor three level statistical
design was used to quantify the influence of PEG 6000 and Clopidogrel on the dissolution profile
of the solid dispersions prepared where PEG 6000 and Clopidogrel were chosen as independent
variables while dissolution rate was chosen as dependent variable. Melt fusion method and solvent
evaporation method were used for the preparation of solid dispersion. Results obtained showed
that there was a significant increase in the dissolution rate of the drug as well as solubility of the
drug in comparison to pure drug. Differential scanning calorimetry. X-ray diffraction and scanning
electron microscopy analysis revealed the formation of solid dispersion of the drug with PEG
6000.
Key words: Clopidogrel, solid dispersion, PEG 6000, Dissolution, Factorial Design
INTRODUCTION
Clopidogrel, methyl (+)--(2-chlorophenyl)-6, 7-dihydrothieno [3, 2-c] pyridine-5(4H)-
acetate, is a non-competitive inhibitor of adenosine diphosphate (ADP) at the platelet receptors. It
is indicated for the early and long term secondary prevention of atherothrombotic events in
patients with acute coronary syndromes (EMEA,2009). Clopidogrel is practically insoluble in
water, which results in poor dissolution and poor bioavailability of the drug. Thus increasing the
aqueous solubility and dissolution of Clopidogrel is of therapeutic importance. A large number of
techniques have been reported over the years to enhance the drug solubility and the dissolution of
the drugs. Some of the examples are formation of inclusion complexes with cyclodextrins
(Loftsson et al.,2004), formation of solid dispersion with hydrophilic carrier (Waghmare et al.,
2008, Dehgan et al.,2006, Rane et al.,2007, Punitha et al.,2010), Micellar drug solubilisation
(Carlota et al., 2005), Dendrimers for drug solubilisation (Gupta et al., 2006), spray drying
techniques (Chauhan et al., 2005), salt synthesis (Bastin et al., 2000), prodrug approach (Banerjee
et al., 2004) and nanoparticle loaded drug molecules (Troy et al., 2006). Solid dispersions are
defined as the dispersion of one or more active ingredients in an inert hydrophilic carrier or matrix
in a solid state (Chiou et al., 1971). Solid dispersion technique has been widely used to incorporate
drug in hydrophilic carriers to enhance the dissolution rate of less water soluble drugs often
leading to increased bioavailability of the drug. In the present study PEG 6000 was used as a
surface adsorbent to formulate solid dispersions of clopidogrel. Polyethylene glycols are addition
polymers of ethylene oxide and water. It has a very high affinity towards water and has been used
 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

extensively for preparing solid dispersions by various research Table 2. Drug Content with Solvent Evaporation method.
groups. S.no. Formulation Code Percentage of Drug
The aim of the present study was to improve the solubility content
1 SF1 73.02±0.57
and dissolution rate of Clopidogrel by formulating a solid
2 SF2 98.15±0.23
dispersion with PEG 6000. Full Factorial experimental designs are
3 SF3 54.7±1.80
one of the best ways to study the effects of different variables on
4 SF4 61.83±0.45
the quality determinant parameters of any formulation. Thus a 5 SF5 93.16±0.40
statistical model was further developed to optimize the solid 6 SF6 81.28±0.28
dispersion formulations. An appropriate statistical model was 7 SF7 91.52±0.08
arrived at and a significant enhanced dissolution rate was observed 8 SF8 94.26±0.48
with the optimized formulation. 9 SF9 97.48±0.22

EXPERIMENTAL Table 3. Drug Content with Melt Fusion Method.

Materials S.no Formulation Percentage of drug
PEG 6000 was obtained from Merck India Ltd. (Mumbai). content
The Clopidogrel was obtained as gift sample from Ranbaxy 1 MF1 95.5±0.45
Laboratories Ltd. (Gurgaon, India). All other chemicals and 2 MF2 82.36±0.37
3 MF3 68.44±0.10
solvents used were of suitable analytical grade.
4 MF4 98.17±0.20
5 MF5 92.23±0.32
Methods
6 FM6 94.16±0.40
Preparation of solid dispersions of clopidogrel with PEG 6000
7 MF7 86.9±0.43
Solid dispersions of clopidogrel in PEG 6000 were 8 MF8 91.81±0.25
prepared using a two factor three level factorial design with 9 MF9 97.55±0.40
clopidogrel and PEG 6000 as independent variables while
maintaining the amount of solvent added as constant. The methods Fusion method
used for preparation of these solid dispersions were solvent A required amount of PEG-6000 melt in a glass container
evaporation method and melt fusion method. on a water bath at about 50-60˚C. A required amount of
clopidogrel dissolved in methanol was added to molten PEG 6000
Solvent evaporation method and mixed thoroughly with a glass rod for 10-15 minutes. Mixture
The required amounts of Clopidogrel and PEG6000 was was cooled rapidly by placing glass container in ice bath (0-4˚ C)
dissolved in methanol and thoroughly mixed for 1 hour over a for 20-25 minutes. A total of nine batches (MF1 to MF9) by fusion
magnetic stirrer. The solvent was then removed at 60˚ C under method were prepared (Table 1).
vacuum inside a Rotary vacuum evaporator until the solid
dispersion was dry. The dried mass was pulverized, passed through Evaluation of solid dispersions
sieve and stored in desiccators over anhydrous CaCl2 until use. A Drug content
total of nine batches (SF1 to SF9) by solvent evaporation method Solid dispersion equivalent to 10 mg of clopidogrel were
were prepared (Table 1). weighed accurately and mixed with suitable quantity of methanol
in a 100ml volumetric flask, volume was made up using distilled
Table 1. Design layout of a Two –Factor, Three- level Factorial Design.
water. Drug content was analyzed at 240 nm using UV
BATCH DRUG POLYMER spectrophotometer (Cary 5000). Each sample was analyzed in
SF1, MF1 -1 -1 triplicate.
SF2, MF2 -1 +1
SF3, MF3 +1 0 Solubility studies
SF4, MF4 +1 -1 The solubility study of the drug and its solid dispersions
SF5, MF5 0 -1 was carried out by taking 20 mg of drug and its solid dispersion
SF6, MF6 -1 0 equivalent to 20 mg of drug in a 25 ml conical flask containing
SF7, MF7 +1 +1 10ml distilled water, volume was made up to 25 ml and shaken on
SF8, MF8 0 +1 a mechanical shaker for 24 hrs at room temperature and analyzed
SF9, MF9 0 0 at 240 nm using UV-Vis spectrophotometer after suitable dilutions.
*Translation of coded units in actual units
In vitro dissolution study
Coded level* -1 0 +1 The dissolution study was carried out using USP
A (DRUG) mg 200 300 400 Dissolution Apparatus 2. (Rotating Paddle type). Accurately
B (PEG 6000) mg 200 400 600 weighed amount of pure drug and solid dispersion were immersed
 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

in a dissolution medium consisting of pH 2.0 HCL buffer RESULTS AND DISCUSSION

maintained at 37◦ C. The dissolution was carried out for 1 hour and
Content of Clopidogrel
aliquot of 5ml was withdrawn at adequate intervals. An equal
The drug content of the prepared formulations of
volume of fresh dissolution medium was replaced (maintained at
Clopidogrel was observed to be varying from 54.7 to 98.15%
the same temperature) in order to keep the total volume constant.
respectively and it was maximum with formulation SF2 and
The filtered samples were suitably diluted, if necessary, and
minimum in formulation SF3 by Solvent evaporation method,
assayed spectrophotometrically at 240nm.
which may be attributed to the fact that insufficient polymer may
not be able to entrap the drug moiety into the solid dispersion
Analytical methods
For Melt Fusion method the drug content of the prepared
Differential scanning calorimetry (DSC)
formulations varied from 68.44 to 98.17 respectively and it was
The DSC thermo grams were recorded using a differential
highest for formulation MF4 and lowest for formulation MF3. It
scanning calorimeter (Q10 TA Instruments, USA). Approximately
was observed that melt fusion method yielded higher drug content
2-5 mg of each sample was placed in an aluminium pan and heated
in formulation with respect to solvent evaporation technique.
from 30 to 300◦ C at a scanning rate of 10˚ C/min under a stream
of nitrogen.
Solubility studies
Total aqueous solubility of the formulations is shown in
Scanning Electron Microscopy (SEM)
Table 4 and 5. In solvent evaporation method the aqueous
The SEM analysis was carried out using a scanning
solubility of drug was found to be maximum formulation SF7
electron microscope (LEO, 435 VP, and UK). Prior to examination
samples were mounted on an aluminium stub and then made (17.57±0.41 g/ml) and minimum with SF4 (10.01±0.15g/ml).
electrically conducting by coating it with a thin layer of gold In melt fusion method the aqueous solubility of drug was observed
(approximately 20nm). The scanning electron microscope was to be maximum with formulation MF7 (17.86±0.32g/ml) and
operated at an acceleration voltage of 15kv. minimum with MF2 (10.09±0.08). Studies show that the maximum
solubility is achieved when the concentration of the polymer and
Powder X -Ray Diffraction Analysis (XRD) the drug is highest. It was observed that the formulation prepared
Powder X-Ray diffraction patterns were recorded using a by melt method showed greater solubility as compared to solvent
Powder X-Ray diffractometer under the following conditions: evaporation method.
target Cu; filter Ni; voltage 35 kV; current 20mA; receiving slit 0.2 Table 4. Solubility of Clopidogrel by solvent evaporation method.
inches. The data were collected in the continuous scan mode using
S.no Formulation Code Solubility (g/ml)
a step size of 0.01◦ at 2θ/s. The scanned range was 5-50◦.
1 SF1 11.67±1.20
2 SF2 11.82± 0.78
Fourier infrared spectroscopy ( FTIR Analysis) 3 SF3 12.25±0.68
The FTIR spectra were recorded on a FTIR multiscope 4 SF4 10.01±0.15
spectrophotometer (Perkin Elmer, UK) equipped with spectrum 5 SF5 10.62±0.33
v3.02 software. KBr pellets of adequate amount of sample were 6 SF6 13.83±0.87
prepared. The spectrum for each sample was recorded over the 450 7 SF7 17.57±0.41
8 SF8 11.9±0.037
to 4000cm-1 spectral region with a resolution of 4 cm-1.
9 SF9 13.84±0.99
10 Drug 9.92± 0.01

Experimental Design Table 5. Solubility of Clopidogrel by Melt fusion method.

A three- level, two factor experimental design as shown in S.no Formulation Code Solubility (g/ml)
Table no.1 describes the proportion in which the independent 1 MF1 11.87±1.26
variables Clopidogrel (A) and PEG 6000 (B) were used in the 2 MF2 10.09±0.08
3 MF3 11.09±0.40
formulation of dispersion granules. The dissolution rate, as % drug 4 MF4 10.56±0.22
release in 15 minutes (D15) and at 60 minutes (D60) was taken as 5 MF5 11.15±0.14
dependent variables. All the nine formulations were prepared and 6 MF6 13.34±0.72
7 MF7 17.86±0.32*
analyzed for dissolution for each method of manufacturing used. 8 MF8 10.76±0.16
Various response surface methodology computations were 9 MF9 14.11±0.75
performed employing Design Expert Software version 8.0.5. 10 Drug 9.92±0.01
Polynomial models including interaction and quadratic terms were
generated for all the response variables using multiple linear In vitro dissolution study
regression analysis approach. In addition contour and surface plots Solvent evaporation method
were also obtained by Design Expert to represent the effect of In vitro dissolution studies of the different formulations of
independent variables graphically. Clopidogrel prepared by solvent evaporation technique, it was
.
 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226
observed that the Solid dispersion of Clopidogrel prepared by this
method showed improved dissolution when compared with pure
drug. The trend observed was an increase in dissolution rate on
increasing the amount of PEG 6000. This could be due to an
increase in the effective surface area over which the drug
distribution increases accompanied by an enhancement in drug
dissolution. Formulation SF7 (Figure 1) showed the maximum
release at the end of 60min with a % drug release of 81.23%. It was
observed that for lower drug concentrations the drug release
increase with increase in polymer concentration.
Fig. 1 Effect of PEG 6000 and Clopidogrel content on % drug
release by Solvent evaporation method.
Fusion method
In vitro dissolution studies of the different formulations of
Clopidogrel prepared by Fusion technique, it was observed that the
Solid dispersion of Clopidogrel prepared by this method showed
improved dissolution when compared with pure drug. The trend
observed was an increase in dissolution rate on increasing the
amount of PEG 6000. Formulation MF7 (Figure 2) showed the
maximum release at the end of 60min with a % drug release of
92.7%.
Fig.2 Effect of PEG 6000 and Clopidogrel content on % drug release by Fusion
method.
Differential scanning calorimetry
In order to study the interaction between Clopidogrel and
PEG 6000 DSC studies were performed on the individual
components. The DSC curve of Clopidogrel showed one
endothermic peak at nearly 183.19˚ C. Pure PEG 6000 showed an
endothermic peak at 63.59˚ C. Thermo grams of solid dispersions
using the solvent evaporation method and fusion method showed
the absence of a Clopidogrel peak, suggesting that Clopidogrel is
either completely soluble in the liquid phase of the polymer or
there is absence of crystalline nature of drug. However, the melting
peak of PEG 6000 in solid dispersions was observed at slightly
lower temperature (56.97˚ C to 59.80˚ C). Absence of an
endothermic peak of drug in solid dispersions has also been
reported by other research groups.
Scanning electron microscopy (SEM)
The surface morphology of Cloidogrel and its solid
dispersion was examined by SEM analysis. Figure 4 shows some
of the selected SEM images of chosen samples. The clopidogrel
crystals appeared with sharp edges and partially agglomerated in
bundles. In solid dispersions by solvent evaporation method and
Fusion method clopidogrel particles were adsorbed on to the
surface of PEG6000 which gives a very rough contour to the
surface this contributed to a higher dissolution rate because of
enhanced effective surface area of clopidogrel. In addition the
SEM images of the batch prepared by melt fusion method showed
greater proportion of PEG 6000 evident from the plate like
structures characteristic of PEG, which could have contributed to
greater dissolution rate of batches prepared by melt fusion method
as compared to solvent evaporation method.
X-Ray diffraction analysis (XRD)
The XRD pattern of Clopidogrel, PEG 6000 and solid
dispersions are given in Figure 5. The XRD pattern of Clopidogrel
showed a large number of intense and sharp peaks indicating its
crystalline nature. Crystallinity was determined by comparing
some representative peak heights in the diffraction patterns of the
solid dispersions prepared with those of the pure drug. It was
observed that Clopidogrel showed sharp peaks at 20.7˚, 23.3˚ and
25.7˚ (2θ) with peak intensities of 5185, 16695 and 5845 cps
respectively. However significant decrease in the peak intensities
of Clopidogrel was observed in the solid dispersions prepared by
using PEG 6000 at the same angles. This indicates a decrease in
the crystallinity of the pure drug.
Fourier transform Infrared Spectroscopy (FTIR) Analysis
IR spectra of binary system of Clopidogrel showed IR
spectra of Clopidogrel and PEG 6000 and no additional peak was
observed in binary systems. IR spectrum of Clopidogrel is
characterized by principal absorption peaks at 3113.16cm-1 (C-H
aromatic), 1748.41.36cm-1 (C=O stretching), 1176.36cm-1 (C-O
stretching). The IR spectra of all solid dispersions show
disappearance of some peaks of Clopidogrel. The peaks at 2720.89
cm-1, 1539.4 cm-1, 1070.12 cm-1 are completely absent. All other
peaks appeared with decreased intensity. Rest of the peaks of
Clopidogrel were found to be smoothened indicating strong
physical interaction of Clopidogrel with PEG 6000. However, no
additional peak was observed in the binary system indicating
absence of any chemical interaction between Clopidogrel and PEG
6000.
Statistical Modelling for Optimization
The statistical evaluation of dependent variables was
performed by Analysis of variance (ANOVA) using Design expert
 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

Fig.3 DSC Spectra of (A) PEG 6000, (B)Clopidogrel , (C) MF2 , (D) SF2.
 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

effect of the factor on the response. The polynomial terms (A2 and
B2) are included to represent the nonlinearity and the interaction
term (AB) shows how the values of the percent drug dissolved in
15 and 60 minutes changes when the two factors are changed
simultaneously. The fitted equations relating the responses D15 and
D60 to the transformed factors for both the fusion method and
solvent evaporation method are shown in equation 2,3,4,5
respectively.

Fusion method

(D15)=60.35-1.03*A+12.69*B+4.91*A*B+5.09*A2 -0.77*B2+
1.45*A2* B+7.12*A*B2 (2)

(D60)=64.32+1.38*A+12.51*B+7.40*A*B+7.66*A2-1.37*B2 (3)

Solvent evaporation method

(D15)=64.04-2.12*A+4.66*B+2.46*A*B+4.03*A2+1.17*B2 (4)
Fig. 4 SEM photographs of (A) Clopidogrel (91X), (B) Solid dispersion by Fusion
method (1000X), (C) Solid dispersion by solvent evaporation method (1000X).
(D60)=65.36-1.59*A+5.27*B+2.97*A*B+5.11*A2+3.14*B2 (5)
Software version 8.0.5. The ANOVA results of the It may be concluded from the equations and the contour
variables on the percent drug dissolved in 15 and 60min from the and 3D plots obtained that the high level of polymer PEG 6000(B)
Solid dispersions prepared by both the Fusion method and Solvent has a more profound effect on the percent drug release at 15 and 60
evaporation method are shown in Table 6 & 7. min than factor A (drug concentration).
Three dimensional response surface plots and two
Table 6. ANOVA results of dependent variables for the fusion method.
dimensional contour plots were obtained from the data with Design
Response Sum of df Mean F value P-value R2 expert version 8.0.5 to estimate the effects of the independent
squares square Prob>F
% DR in 1444.87 7 206.41 665.10 <0.0001 0.9989 variables (A and B) on each response, as depicted in Fig.7 and 8.
15min(D15 )
% DR in 1340 5 268 55.21 <0.0001 0.9753
60min(D60 )
CONCLUSION
The enhancement of solubility of a poorly soluble drug
Table7. ANOVA-influence of dependent variables for the solvent evaporation
method. Clopidogrel from its solid dispersion with PEG 6000 has been
investigated. The solid state properties of the binary system were
Response Sum of df Mean F P-value R2
squares square value Prob>F characterized by powder XRD, DSC, SEM and FTIR studies
% DR in 250.57 5 50.01 25.90 0.0002 0.9487 showed decrease in the crystallinity of the drug in the solid
15min(D15)
% DR in 373.20 5 74.64 17.40 0.0008 0.9255 dispersions prepared. The study showed that the dissolution rate of
60min(D60) the drug can be enhanced to a great extent by solid dispersion
technique using both the solvent evaporation and Fusion method.
The effects of independent variables upon the responses
were modelled using the following general form of the equation 1.
ACKNOWLEDGEMENT

Y= b0+b1A+b2B+b3AB+b4A2 +b5B2 +b6AB 2+b7A2B (1)

We are thankful to the Incharge, Central Instrumentation
Where Y is the predicted response, b0 is the arithmetic mean
Laboratory GJUS&T, Hisar for DSC and UV spectrophotometric
response of 13 runs where b1 and b2 are the estimated coefficients
studies. Department of Anatomy, All India Institute of Medical
for the factors A and B respectively. The main effects (A and B)
Sciences, New Delhi for SEM studies. Head, Department of
represent the average result of changing one factor at a time from
Physics GJUS&T, Hisar for XRD studies. We are also thankful to
its low to high value.
RANBAXY LABORATORIES Gurgaon, INDIA for providing
Coefficients with one factor indicate the effect of that
gift sample of CLOPIDOGREL.
particular factor on the desired response, while the coefficients
with more than one factor and those with second-order terms
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 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

Fig.5 XRD Spectra of (A) Clopidogrel, (B) PEG 6000, (C) Formulation.
Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

Fig.6 FTIR Spectra of (A) Clopidogrel, Formulation (B) MF5 & (C) SF2.
 Journal of Applied Pharmaceutical Science 01 (08); 2011: 217-226

Fig.7 Contour and surface plots showing % drug release in 15 minutes and 60 minutes of solid dispersions as a function of PEG 6000 and Clopidogrel for Solvent
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Fig.8 Contour and surface plots showing % drug release in 15 minutes and 60 minutes of solid dispersions as a function of PEG 6000 and Clopidogrel for
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